正常人骨髓成纤维样基质细胞对HL-60/VCR耐药细胞增殖的影响  

Effects of normal human bone marrow fibroblastoid stromal cell line on the proliferation of HL-60/VCR multidrue resistant cell line

作　　者：梁蓉[1] 黄高昇[1] 王哲[1] 冯骥良[1] 张伟平[1] 杨国嵘[1] 郭英[1] 王娟红[1] 

机构地区：[1]第四军医大学基础部病理教研室,西安710033

出　　处：《肿瘤》2004年第5期429-432,共4页Tumor

基　　金：国家自然科学基金资助项目(编号:30300141)

摘　　要：目的 观察正常人骨髓成纤维样细胞系HFCL对白血病多药耐药细胞HL-60／VCR增殖和分化的影响。方法 采用四唑氮蓝(MTT)法进行HL-60／VCR细胞药物敏感实验。建立HL-60／VCR细胞和HFCL细胞共培养体系,苔盼蓝拒染法测定生长曲线;硝基四氮唑蓝(NBT)确定细胞分化;流式细胞仪检测细胞周期和CD11b、CD13、CD14、CD33细胞表面抗原进一步鉴定细胞分化;Western blot检测增殖细胞核抗原(Proliferating Cell Nuclear Antigen,PCNA)和P糖蛋白(P-glycoprotein,P-gp)。结果 HL-60／VCR细胞对多种药物耐药。与HFCL细胞共培养后,HL-60／VCR细胞生长受抑,且与HFCL细胞直接接触组的抑制作用>用transwell组。同时发现HL-60／VCR细胞与HFCL细胞共培养后,G1期细胞增多,S期细胞减少;同时CD11b和CD14表达增高,CD13和CD33变化不大;且NBT阳性细胞轻度增多。Western blot检测结果显示,PCNA表达下调,以直接接触组为甚。但是P-gp表达无变化。结论 正常人骨髓成纤维样细胞HFCL能抑制白血病MDR细胞HL-60／VCR的增殖,抑制PCNA的表达,出现G1期阻滞,并部分向单核细胞分化。Objective To investegate the effects of normal human bone marrow fibroblastoid stromal cell line(HFCL) on the proliferation and differentiation of acute myeloid leukemia multidrug resistant (MDR) cell line HL-60/VCR. Methods Drug sensitive test was shown by MTT assay. Establishing coculture system of leukemia MDR cell line HL-60/VCR and HFCL cell, growth curves were detected by cell counting. Cell differentiation was determined by morphologic observation, ability of NBT cells and flow cytometric detection of the expression of CD11b,CD14,CD13 and CD33. Flow cytometry was also performed to test the change of cell cycle. Western blot was used to study the expression of proliferating cell nuclear antigen (PCNA) and P glycoprotein(P-gp). Results HL-60/VCR cells were multidrug resistant. Compared with HL-60/VCR cells without HFCL cells, the proliferation of HL-60/VCR cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited. And NBT positive cells increased slightly. The percentage of G1 phase cells of HL-60/VCR cells with HFCL cells was higher than that without HFCL cells, and the percentage of S phase cells was lower. Meanwhile the expression of CD11b and CD14 increased, however, the expression of CD13 and CD33 didn't change. The expression of PCNA in HL-60/VCR cells with HFCL cells was lower than HL-60/VCR cells without HFCL cells. The expression of P-gp of HL60/VCR cells didn't change in the condition of coculture. Conclusion The normal bone marrow fibroblastoid stromal cells HFCL could inhibit the proliferation and progress of cell cycle of acute myeloid leukemia MDR cells HL-60/VCR, and also could induce the differentiation of part of HL-60/VCR cell into monocyte.

关 键 词：骨髓成纤维样基质细胞HFCL HL-60/VCR细胞 多药耐药 细胞增殖 细胞分化 

分 类 号：R733.7[医药卫生—肿瘤] R73-362[医药卫生—临床医学]